Recombinant high molecular weight-glutenin subunit-specific IgE detection is useful in identifying wheat-dependent exercise-induced anaphylaxis complementary to recombinant omega-5 gliadin-specific IgE test
Recombinant high molecular weight-glutenin subunit-specific IgE detection is useful in identifying wheat-dependent exercise-induced anaphylaxis complementary to recombinant omega-5 gliadin-specific IgE test
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DOI：
10.1111/j.1365-2222.2012.04039.x
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发表时间：
2012-08-01
影响因子：
6.1
通讯作者：
Morita, E.
中科院分区：
文献类型：
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作者：
Takahashi, H.;Matsuo, H.;Morita, E.
研究背景小麦依赖性运动过敏反应（Wheat-dependent exercise-induced allergysaccharide，WDEIA）是一种特殊形式的食物过敏反应，通常由摄入小麦制品后运动引起。我们确定了小麦ω-5麦醇溶蛋白和高分子量麦谷蛋白亚基（HMW-麦谷蛋白）作为WDEIA的主要过敏原，并阐明了血清IgE结合这些过敏原的合成表位肽的同时检测识别超过90%的WDEIA患者。目的构建重组HMW-谷蛋白（HMW-glutenin）荧光酶免疫分析（CAP-FEIA），并评价其在WDEIA患者诊断中的应用价值。大肠杆菌中表达，用组氨酸标签亲和层析柱纯化。48名WDEIA患者血清中的小麦、谷蛋白、重组ω-5麦醇溶蛋白、HMW-麦谷蛋白表位肽、天然和重组HMW-麦谷蛋白特异性IgE，应用CAP-FEIA法对16例无小麦过敏反应的AD患者和12例健康对照者进行了过敏原检测。天然高分子量麦谷蛋白试验阳性率为12.5%，重组高分子量麦谷蛋白试验阳性率为12.5%。这些结果表明天然HMW-谷蛋白测试具有低特异性。重组高分子量谷蛋白检测IgE的敏感性为16.7%，特异性为92.9%。这些结果与利用高分子量麦谷蛋白表位肽获得的结果一致。敏感性和特异性分别为93.8%和92.9%，结论和临床相关性我们证明了重组HMW-麦谷蛋白在稳定性和特异性方面对于CAP-FEIA系统是最好的，并且证实了当与CAP-FEIA组合时，检测针对重组HMW-麦谷蛋白的特异性IgE对于WDEIA的诊断是有用的（重组ω-5麦醇溶蛋白）测试。
Background Wheat-dependent exercise-induced anaphylaxis (WDEIA) is a special form of food allergy typically induced by exercise after ingestion of wheat products. We identified wheat omega-5 gliadin and high molecular weight-glutenin subunit (HMW-glutenin) as major allergens for WDEIA and clarified that simultaneous detection of serum IgE binding to synthetic epitope peptides of these allergens identifies more than 90% of WDEIA patients. However, the short synthetic peptides are not suitable for CAP-fluorescent enzyme-immunoassay (CAP-FEIA), which is widely utilized for detecting allergen-specific IgE.Objective In this study, we constructed a CAP-FEIA with recombinant HMW-glutenin, and evaluated its usefulness in identifying the patients with WDEIA.Methods Recombinant HMW-glutenin was expressed as histidine-tag protein in E. coli and purified by histidine-tag affinity column. Wheat, gluten, recombinant omega-5 gliadin, epitope peptide of HMW-glutenin, native and recombinant HMW-glutenin specific IgE in the sera from 48 patients with WDEIA, 16 patients with atopic dermatitis (AD) who had no immediate allergic reaction after wheat ingestion and 12 healthy controls were determined by using CAP-FEIA method.Results In 16 AD patients without wheat allergy 12 of them (75%) had positive results for native HMW-glutenin test in contrast to epitope peptide of HMW-glutenin (12.5%) and recombinant HMW-glutenin test (12.5%). These results indicate the native HMW-glutenin test has low specificity. Sensitivity and specificity of the IgE test with recombinant HMW-glutenin were 16.7% and 92.9%. These are well compatible with results obtained by using epitope peptide of HMW-glutenin. However, sensitivity and specificity reached to 93.8% and 92.9%, when the test was combined to the test with recombinant omega-5 gliadin.Conclusions and Clinical Relevance We demonstrated that recombinant HMW-glutenin is best for CAP-FEIA system in point of stability and specificity and confirmed that detection of specific IgE against recombinant HMW-glutenin is useful for diagnosis of WDEIA when combined with the CAP-FEIA (recombinant omega-5 gliadin) test.